ObjectiveTo evaluate the antitumor activity of Periplaneta americana extract（PAE） against human-derived lung adenocarcinoma organoids（LUAD-PDOs） and to elucidate its potential mechanism based on transcriptomics. MethodsFresh tumor and adjacent normal tissues from patients with LUAD were collected to construct LUAD-PDOs and normal lung organoid（Nor-PDOs） models using 3D organoid culture technology. The effective intervention concentration of PAE was determined using the cell counting kit-8（CCK-8） assay. Experimental groups included the model group（LUAD-PDOs）， normal group， model administration group（LUAD-PDOs+PAE）， and normal administration group（Nor-PDOs+PAE）. Hematoxylin-eosin（HE） staining was used to observe the pathological structures of PDOs， immunohistochemistry（IHC） was performed to detect the expressions of the proliferation marker Ki-67 and lung adenocarcinoma differentiation markers cytokeratin-7（CK-7） and Napsin A， TUNEL staining was applied to detect cell apoptosis. RNA sequencing（RNA-Seq） was conducted to identify differentially expressed genes（DEGs）， followed by Gene Ontology（GO）， Kyoto Encyclopedia of Genes and Genomes（KEGG）， and Gene Set Enrichment Analysis（GSEA）， alongside protein-protein interaction（PPI） network analysis to screen core mechanisms. Finally， key targets were validated by integrating external database analysis with immunofluorescence（IF）. ResultsNor-PDOs and LUAD-PDOs that highly recapitulated the pathological characteristics of the primary tissues were successfully established. The CCK-8 assay determined that the effective intervention concentration of PAE was 16 g·L^-1. Morphological observation showed that Nor-PDOs exhibited lumen-forming structures， whereas LUAD-PDOs displayed dense， solid structures. CCK-8 and TUNEL assays revealed that， compared with the model group， PAE intervention inhibited the proliferation of LUAD-PDOs and promoted apoptosis in LUAD cells， while showing no significant effect on the viability of Nor-PDOs. Transcriptomic analysis identified 719 DEGs that were significantly reversed after PAE intervention（347 up-regulated and 372 down-regulated）（P<0.05）. GO enrichment analysis indicated that DEGs in the model administration group were significantly enriched in biological processes related to cell cycle regulation compared to the model group. KEGG pathway analysis revealed that PAE affected pathways related to proliferation and metabolism， including pathways in cancer and the p53 signaling pathway. GSEA further confirmed that PAE significantly enhanced the activity of the p53 signaling pathway（P<0.05）. PPI network analysis indicated that breast cancer type 1 susceptibility protein（BRCA1） and checkpoint kinase 1（CHEK1） were the core down-regulated targets in the p53 pathway. IF verified the high expression of BRCA1 and CHEK1 in LUAD-PDOs and their significant downregulation after PAE intervention（P<0.05）. Furthermore， survival analysis based on The Cancer Genome Atlas（TCGA） database indicated that low expression of BRCA1 and CHEK1 was significantly associated with prolonged overall survival in patients with LUAD（P<0.05）. ConclusionPAE effectively inhibits proliferation of LUAD-PDOs and promotes their apoptosis， its anti-tumor mechanism is potentially associated with the activation of the p53 signaling pathway， with BRCA1 and CHEK1 genes likely serving as key downstream targets for the effects of PAE.

Antibody-drug conjugates (ADCs) have emerged as a standard treatment modality for previously treated driver gene-positive advanced non-small cell lung cancer (NSCLC). Sacituzumab tirumotecan (sac-TMT), a novel TROP-2-targeting ADC, has been the focus of the phase Ⅲ clinical trial OptiTROP-Lung04. This trial demonstrated, for the first time, that single-agent sac-TMT significantly improves progression-free survival and overall survival compared with conventional platinum-based chemotherapy in patients with epidermal growth factor receptor (EGFR) mutant advanced NSCLC following resistance to EGFR tyrosine kinase inhibitors. This review combines existing consensus from literature with the latest research evidence to systematically analyze OptiTROP-Lung04 from multiple perspectives, including molecular mechanisms, study design, clinical significance, and safety management. Furthermore, it explores future development directions in an effort to provide new insights into precision therapy for advanced NSCLC.

Objective To analyze the characteristics and drug resistance of pathogenic bacteria isolated from patients undergoing solid organ transplantation(SOT)at West China Hospital,Sichuan University in re-cent years,in order to provide a basis for empirical anti infective treatment after SOT surgery.Methods A retrospective analysis was conducted on the isolation of major pathogens and their resistance to common anti-biotics from various specimens collected from patients undergoing kidney transplantation(KT),liver trans-plantation(LTx),and lung transplantation(LT)at West China Hospital,Sichuan University from 2017 to 2022.Results A total of 1 077 non-duplicate strains of pathogens were isolated from the samples of patients with infections after SOT surgery during the 6-year period,of which approximately 74.8％(806/1 077)were Gram negative bacteria and 25.2％(271/1 077)were Gram positive bacteria.There were differences in the distribution of pathogenic bacteria among different types of SOT groups and different specimens.Compared to E.coli isolated from urine specimens,the strains of E.coli isolated from non-urinary specimens exhibited a higher resistance rate to common antimicrobial drugs(P＜0.05).The resistance rate of E.coli to β-lactam/β-lactamase inhibitor combinations(cefoperazone/sulbactam and piperacillin/tazobactam)in the LTx group was significantly higher than that in the KT group(P＜0.05).The overall proportion of multidrug-resistant bac-teria after SOT surgery was 11.3％(122/1 077).The proportion of carbapenem resistant Acinetobacter bau-mannii and carbapenem resistant Pseudomonas aeruginosa among the same group and type of pathogens in the LTx group(93.8％,37.5％)was significantly higher than that in the KT group(55.8％,9.2％).Conclusion The specimen types and strain distribution of pathogenic bacteria after different types of SOT surgery are different.The same pathogenic bacteria have different antibiotic resistance among different types of SOT groups and specimens.Therefore,it is necessary to strengthen the pathogen examination after different types of SOT and optimize the anti infection treatment plan related to transplantation based on drug sensitivity re-sults.

Objective: To establish a mouse model of lung injury induced by self-antigens and explore its pathological mechanisms to provide a reliable animal model for studying the pathogenesis of rheumatoid arthritis-associated interstitial lung disease(RA-ILD). Methods: The mice were injected intradermal with antigenemulsion made of complete freund′s adjuvant(CFA) and lung tissue protein, and the emulsion prepared with incomplete Freund′s adjuvant(IFA) was used to enhance immunity. HE staining was used to observe the histopathological changes in the mice. Masson staining was used to detect pulmonary fibrosis. The mRNA levels of rheumatoid factor(RF), krebs von den lungen-6(KL-6) and surfactant protein D(SP-D) were detected by qPCR. The levels of ACPA, IgG1, IgG2a and IgG3 antibodies were detected by ELISA. The changes of inflammatory cells and α-smooth muscle actin(α-SMA)expression in lung tissue were detected by immunofluorescence staining. The protein expression levels of Collagen I and α-SMA were detected by Western blot. The changes of immune cells were detected by flow cytometry. Results: HE staining showed that inflammatory cell infiltration increased and tissue structure changed significantly in lung tissue after the model was established by self-lung tissue antigen. Masson staining showed increased collagen deposition in lung tissue of model mice. qPCR tests revealed elevated mRNA levels of RF, KL-6 and SP-D. ELISA tests revealed elevated levels of ACPA, IgG1 and IgG3 antibodies. Immunofluorescence results showed that monocytes and T cells increased, and α-SMA expression increased in the model group. Western blot results showed increased protein expressions of Collagen Ⅰ and α-SMA. The flow cytometry results showed an increase in T cells and monocytes in the lung tissue. Conclusion The mouse model of lung injury induced by self-antigens is successfully established, and T cells and monocytes may be involved in the occurrence and progression of the disease.

Objective: To explore the role of tumour necrosis factor-α(TNF-α) in splenic infection of mice by Leishmania major. Methods: To establish an infection model, promastigotes of Leishmania were injected intradermally into the right hind foot of mice. The thickness of the footpad and body weight were measured to monitor the infection. Histological changes in the spleen after infection were observed by HE staining. Changes in lymphocytes and monocytes in the spleen were detected by flow cytometry. The expression level of arginase-1(Arg-1) and inducible nitric oxide synthase(iNOS) in the spleen was determined by indirect immunofluorescence. The effect of TNF-α on macrophage infection with Leishmania was evaluated in vitro. Results: Compared to B6.WT mice, the spleens of B6.TNF-/- mice showed significant enlargement 42 days post-infection, with structural disruption. Various cells infiltrated and were dispersed throughout the entire spleen. Flow cytometry results indicated that after infection with Leishmania, there was no significant change in the proportions of T cells and B cells in the spleens of the mice, while CD11b monocytic cells significantly increased. Immunofluorescence results revealed that the M2 macrophage/monocyte marker Arg-1 was highly expressed in the spleens of B6.TNF-/- mice(P < 0. 05) . The expression of iNOS in the spleens of B6. WT mice was relatively strong (P < 0. 05) . In vitro studies found that the absence or inhibition of TNF⁃α significantly increased the infection of Leishmania by peritoneal macrophages ( P <0. 01), while the addition of TNF-a markedly inhibited this infection (P <0. 01). Conclusion The splenic infec-tion in B6. TNF mice following subcutaneous inoculation of Leishmania in the hind footpad may be associatedwith the absence of TNF-a, which leads to M2-type differentiation of macrophages and reduced nitric oxide producton.

Mutations in ion channel genes have long been implicated in a spectrum of epilepsy syndromes. However, therapeutic decision-making is relatively complex for epilepsies associated with channelopathy. Therefore, in the present study, we used a patient-derived organoid model with a novel SCN2A mutation (p.E512K) to investigate the potential of utilizing such a model as a platform for preclinical testing of anti-seizure compounds. The electrophysiological properties of the variant Nav1.2 exhibited gain-of-function effects with increased current amplitude and premature activation. Immunofluorescence staining of patient-derived cortical organoids (COs) displayed normal neurodevelopment. Multielectrode array (MEA) recordings of patient-derived COs showed hyperexcitability with increased spiking and remarkable network bursts. Moreover, the application of patient-derived COs for preclinical drug testing using the MEA showed that they exhibit differential responses to various anti-seizure drugs and respond well to carbamazepine. Our results demonstrate that the individualized organoids have the potential to serve as a platform for preclinical pharmacological assessment.
Organoids/physiology* ; NAV1.2 Voltage-Gated Sodium Channel/genetics* ; Humans ; Anticonvulsants/pharmacology* ; Epilepsy/drug therapy* ; Mutation ; Cerebral Cortex/drug effects* ; Action Potentials/drug effects* ; Carbamazepine/pharmacology*

Objective To establish a sandwich enzyme-linked immunosorbent assay (ELISA) method for the quantitative detection of Chromogranin A (CHGA) in saliva, and to explore its preliminary application in the testing of saliva samples. Methods Recombinant human CHGA protein was used to immunize BALB/c mice, and monoclonal antibodies (mAbs) were prepared and screened using conventional hybridoma technology. A double-antibody sandwich ELISA detection method was constructed, and the matrix effect of saliva samples was optimized. This method was then applied to detect the concentration of CHGA in the saliva of stressed individuals. Results Twenty-one stable hybridoma cell lines secreting high affinity anti-human CHGA antibodies were obtained. A pair of detection antibodies with the best effect was selected, and the optimal coating concentration was determined to be 10 μg/mL, with the optimal dilution of detection antibodies being 1:32 000. The accuracy and reproducibility of this method were verified, with both intra-batch and inter-batch variation coefficients less than 15×, and the recovery rate between 80× and 120×. The matrix effect was further optimized to make it suitable for saliva sample detection. Saliva samples from individuals in different stress states were collected, and the CHGA levels were detected using the method established in this study, indicating its potential to reflect the intensity of stress. Conclusion A reliable saliva CHGA ELISA detection method has been successfully established, and its potential as a biomarker in stress-related research has been preliminarily explored.
Saliva/metabolism* ; Enzyme-Linked Immunosorbent Assay/methods* ; Humans ; Animals ; Mice, Inbred BALB C ; Mice ; Chromogranin A/immunology* ; Antibodies, Monoclonal/immunology* ; Female ; Male ; Reproducibility of Results ; Adult

Advances in targeted therapy and immunotherapy have significantly improved clinical outcomes for patients with advanced non-small cell lung cancer (NSCLC), reshaping treatment paradigms. However, most patients ultimately face drug resistance, with limited options for subsequent therapies and suboptimal treatment efficacy, presenting a prominent challenge in current clinical practice. Antibody-drug conjugates (ADCs), characterized by high efficacy and favorable safety profiles, have emerged as a promising therapeutic frontier in recent years. This systematic review provides a comprehensive overview of the latest advancements in ADCs-based therapies for lung cancer, alongside discussions of the prevailing challenges in this rapidly evolving domain.
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Humans ; Carcinoma, Non-Small-Cell Lung/immunology* ; Immunoconjugates/therapeutic use* ; Lung Neoplasms/immunology* ; Antineoplastic Agents/therapeutic use* ; Immunotherapy

Objective:To investigate the characteristics of clinical manifestations and parameters of 24 hour multichannel intraluminal impedance and pH monitoring (24 h MII-pH) in patients with low mean nocturnal baseline impedance (MNBI) of proximal esophagus.Methods:From November 4, 2014 to February 18, 2024, 308 patients who underwent 24 h MII-pH at Peking University First Hospital due to typical gastroesophageal reflux disease symptoms and(or) extra-esophageal symptoms were retrospectively enrolled. MNBI at 17 or 15 cm above the lower esophageal sphincter (LES) < 2 292 Ω was defined as low proximal esophageal impedance (LPEI), both MNBI at 17 and 15 cm above the LES ≥ 2 292 Ω was defined as normal proximal esophageal impedance (NPEI). The 24 h MII-pH parameters were compared between patients with LPEI and patients with NPEI, as well as the incidence of extra-esophageal symptoms. And the 24 h MII-pH parameters were compared between patients with and without extra-esophageal symptoms. Independent sample t-test, Mann-Whitney U test and chi-square test were used for statistical analysis. Results:Among the 308 patients, 71 patients with LPEI, 236 patients with NPEI, and 1 patient was excluded due to missing the 24 h MII-pH data; and there were 215 patients with extra-esophageal symptoms and 93 without extra-esophageal symptoms. The proportion of extra-esophageal symptoms in the LPEI patients was higher than that in the NPEI patients(81.7% (58/71) vs. 66.1% (156/236)), the times of postprandial total reflux, postprandial acid reflux detected by impedance, proximal total reflux, and proximal acid reflux in the LPEI patients were more than those in the NPEI patients (22.5 (22.8) vs. 17.0 (19.0), 10.5 (13.3) vs. 7.0 (13.0), 9.0 (12.0) vs. 5.0 (11.0), 5.0 (10.0) vs. 3.0 (7.0)), and the differences were statistically significant( χ2=6.28, Z=-1.99, -2.06, -2.26 and -2.44; all P<0.05). The times of weak acidic reflux at supine position, proximal total reflux, proximal acid reflux, and proximal non-acid reflux of the patients with extra-esophageal symptoms were more than those in patients without extra-esophageal symptoms (2.0(5.0) vs. 1.0(4.0), 6.0(13.0) vs. 4.0(10.0), 4.0(10.0) vs. 3.0(7.0), 2.0(4.0) vs. 1.0(3.0)), the MNBI at 15 cm above the LES in patients with extra-esophageal symptoms was lower than that in patients without extra-esophageal symptoms ((3 222.4±1 018.7) Ω vs. (3 512.3±1 032.1) Ω), and the differences were statistically significant ( Z=-2.32, -2.25, -2.00 and -2.13, t=-2.28; all P<0.05). Conclusions:LPEI patients have higher proportion of extra-esophageal symptoms, more times of proximal and postprandial acidic reflux. The proximal esophageal impedance and proximal reflux parameters should be emphasized in the diagnosis and treatment of patients with extra-esophageal symptoms.

Objective:This study utilizes data from the 2021 Chinese Longitudinal Healthy Longevity and Happy Family Survey(CLHLS-HF)to examine the current status of Long-Term Care Insurance(LTCI)implementation and to identify the factors influencing whether disabled elderly individuals receive formal care services.The study aims to provide policy recommendations to enhance the effectiveness and equity of the system.Methods:In this cross-sectional study, a sample of 1 447 older participants with dependency, residing in LTCI pilot areas and meeting the inclusion criteria from the 2021 CLHLS-HF, was selected.Chi-square tests and binary logistic regression analyses were employed to explore the factors influencing the receipt of formal care by dependent older individuals.Results:Among the 1 447 participants, there were 496 males with an average age of 92 years(SD 9)and 951 females with an average age of 95 years(SD 9). Of these, 701 received formal care.The logistic regression analysis revealed that factors influencing the receipt of formal care included urban residence( OR=2.237, 95% CI: 1.675-2.987, P<0.001), residing in the eastern region( OR=2.907, 95% CI: 1.747-4.837, P<0.001), living in the western region( OR=3.132, 95% CI: 1.816-5.501, P<0.001), having no children( OR=2.478, 95% CI: 1.108-5.540, P=0.027), and the degree of disability, with severe disability being more likely to receive care compared to mild( OR=0.497, 95% CI: 0.388-0.637, P<0.001)and moderate disabilities( OR=0.589, 95% CI: 0.433-0.801, P=0.001). Conclusions:Dependent older individuals in the eastern and western regions, particularly those without children or with severe disabilities, are more likely to receive formal care through the LTCI system.However, there are substantial inequities in LTCI coverage among individuals with varying degrees of disability.To enhance the effectiveness of the LTCI system, greater efforts should be directed towards economically disadvantaged regions and older individuals with mild to moderate disabilities, thereby ensuring better protection for the disabled population.